GA/T 1963-2021 in English

VALID

Forensic science -Methods for SSR detection of species identificationof Papaver somniferum L.- Capillary electrophoresis and fluorescence method

Issued on:2021-10-14
Implemented on:2022-05-01
File Format:PDF
Delivery:Via email within 1~3 business days

Price(USD)： $90.00
$88.00

Standard No:	GA/T 1963-2021
Document status:	VALID
Title in English:	Forensic science -Methods for SSR detection of species identificationof Papaver somniferum L.- Capillary electrophoresis and fluorescence method
Title in Chinese:	法庭科学罂粟种属SSR标记检测毛细管电泳荧光检测法
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Issued on:	2021-10-14
Implemented on:	2022-05-01
Professional Classification:	GA-Security
Related Keywords:	capillary electrophoresis fluorescence detection technology platform elementsthis standard methodtraditional morphological methodchemical detection method detection ssr detection l.- capillary electrophoresis fluorescence method introduction analysis
Related Topics:	capillary electrophoresis poppy Fluorescence detection detection method Determination of Inorganic by Capillary Electrophoresis Used Fluorescence Detectors capillary electrophoresis ssr Fluorescence detection standard multiple fluorescent labels marker detection Fluorescence detection Fluorescence detection equipment ssr+ What are the second-hand fluorescence detectors? Fluorescence detector peak Beike Testing detection science fluorescence detection ssr detection fluorescence detection ssr detection Metal fluorescence detection Mark detection system Fluorescence detector cell species Fluorescence Detector Detector fluorescent labeling Capillary Electrophoresis+ forensic science Optical detection model Fluorescence detection food detection Detection of autofluorescence fluorescent labeling method Photodetection fluorescent labeling Type of electrophoresis capillary electrophoresis labeled fluorescent method Type of electrophoresis forensic science Fluorescence detection plate Optical detection Lai detection Fluorescence detector series capillary electrophoresis method Fluorescence detection polycapillary detector capillary detection labeled fluorescent detection court Fluorescence Used Detectors None detected Standard method of capillary electrophoresis Internal standard method detection Optical detection model capillary electrophoresis fluorescence Three kinds of electrophoresis ssr batch detection Labeled and unlabeled detection Fluorescence and UV detectors Detection of far-infrared fluorescent labels Fluorescence electrophoresis Metal fluorescence detection Quantitative detection by fluorescence method Deuteration detection Fluorescent detection Fluorescent color detection Chemiluminescent detection capillary electrophoresis electrophoresis Fluorescence detection five-point detection plate Fluorescent Labels in Flow Cytometry fluorescence capillary analysis science mark flow fluorescent labeling Detection of fluorescent labels Cytofluorescence assay capillary fluorescence technique Fluorescence labeling cytoskeleton Luciferase labeling Capillary sequencer detection GA/T 1773.2-2021 GA T 1788.3-2021 GA 1800.2—2021 The latest capillary testing standards Detection of electrophoresis parts Fluorescence detection accepted Histamine fluorescence detection method Local standard fluorescence detection Fluorescent labeling method of microtubules Luciferase method for detecting bacteria Fluorescein detection method Spectral detection method types international forensic science Commonly used fluorescent labeling methods for organelles Cell organelle fluorescent labeling method Specific methods for detecting fluorescent markers using microplate readers Metal fluorescence detection charges Which food testing standards use fluorescence detectors? Capillary electrophoresis detection Fluorescence and other detection methods Cell membrane damage fluorescence detection method Disadvantages of capillary electrophoresis method detection Advantages of capillary electrophoresis method detection Hair heavy metal detection Draft standard for capillary electrophoresis method ga/t1921-2021 Labeling methods for fluorescent nanolabels Cell calcium fluorescence detection method Are there qualitative standards for fluorescence detectors? Electrophoresis salt test Isoelectric focusing electrophoresis detection method

Introduction

Analysis of the technical framework of the standard

This standard is based on the capillary electrophoresis fluorescence detection technology platform to establish a molecular identification system for opium poppy and other plants of the same genus. The core adopts a composite detection scheme of 5 SSR loci (P136, BP13, BP12, P85, BP14), and its specificity is derived from the conservative microsatellite sequence characteristics in the opium poppy genome.

Technical elements	This standard method	Traditional morphological method	Chemical detection method
Detection object	DNA molecular characteristics	Plant morphology	Morphine content
Sensitivity	0.5-1.0ng DNA	Intact organs required	≥seedling stage
Specificity	Joint determination of 5 loci	Highly subjective	Easily affected by processing

Key operating points

1. Primer system design

Five pairs of fluorescently labeled primers were used with a FAM/HEX dual-color labeling system, of which the P136 locus (GAAA repeat sequence) had the highest detection sensitivity (0.3 μmol/L concentration). Special attention should be paid to the adjustment of the primer dosage (0.6 μmol/L) of the BP12 locus during composite amplification.

2. Optimization of amplification conditions

A three-step annealing program was used (59℃→60℃ gradient):
- Initial denaturation: 95℃ 11min
- 28 cycles: 94℃ 30s → gradient annealing→72℃ 2min
- Final extension: 60℃ 60min

Analysis of implementation difficulties

Case: Handling abnormal electrophoresis results

When a laboratory used the 3500xL Genetic Analyzer for detection, the BP14 locus fragment shift (197bp→195bp) occurred. After investigation, the following results were found:
1) Batch differences in molecular weight internal standards
2) Insufficient formamide purity (needed to be ≥99.5%)
Solution: Run positive controls simultaneously and calibrate system errors.

Technology Development History

Compared with the general DNA extraction method of GA/T 1160-2014, the innovations of this standard are:
1) Establishment of opium poppy-specific SSR marker library
2) Development of a composite amplification system (saving 60% of detection time)
3) Clarification of the fragment length determination threshold (200-10000 RFU)

More samples: DL/T 5154-2012 JGJ 94-2008 NB/T 47013.2-2015

Sample only — not a preview of GA/T 1963-2021

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